造血細胞の遺伝子操作

2000 Fiscal Year Annual Research Report

• Project/Area Number: 10181105
• Research Institution: Jichi Medical University
• Principal Investigator: 小澤 敬也 自治医科大学, 医学部, 教授 (30137707)
• Co-Investigator(Kenkyū-buntansha): 吉田 進昭 東京大学, 医科研, 教授 (10250341) ; 北村 俊雄 東京大学, 医科研, 客員教授 (20282527) ; 谷 憲三朗 東京大学, 医科研, 助教授 (00183864) ; 竹田 潤二 大阪大学, 医学部, 教授 (50163407) ; 生田 宏一 京都大学, 医学研究科, 助教授 (90193177)
• Keywords: 遺伝子治療 / 造血幹細胞 / 選択的増幅遺伝子 / コモンマーモセット / レトロウイルスベクター / 突然変異マウス / サイトカインレセプター / トランスポゾンシステム

Research Abstract

体細胞あるいは生殖系列細胞の遺伝子操作を軸とした造血システムの研究を行った。
[造血幹細胞の直接的遺伝子操作]
1.遺伝子導入造血幹細胞の選択的増幅システムの開発(小澤):これまで開発を進めてきた選択的増幅遺伝子の評価をマウスやサルの系で行い、体内で機能することを確認した。
2.小型霊長類造血幹細胞の特性解明と遺伝子導入(谷):GFP発現HIV(VSV)シュードタイプウィルスベクターを用いて、ヒトならびにコモンマーモセットの造血幹細胞へ遺伝子導入することができた。
3.造血細胞用レトロウイルスベクターシステムの開発(北村):エコトロピックウイルスのパッケージング細胞PLAT-Eと造血幹細胞用ベクターpMYの組み合わせで高力価のレトロウイルスベクターを産生することができた。
[発生工学的アプローチによる造血システムの解析]
1.発生工学的手法を用いた造血系システムの解析(吉田):乳糜腹水、リンパ浮腫を特徴とし、リンパ管発生異常と考えられる突然変異マウスの解析を行った。
2.IL-7Rのリンパ球増殖分化シグナル(生田):IL-7レセプターのシグナル伝達分子のSTAT5が、5'Jglプロモーター領域に転写共役因子をリクルートし、ヒストンのアセチル化を介してTCRγ遺伝子座のaccessibilityを上昇させ、germline転写とV-J組換えを誘導することを示した。
3.マウストランスポゾンシステムの開発(竹田):GFP遺伝子を指標にマウス個体での遺伝子トランスポジションを観察した。高頻度にトランスポジションが確認できたので、今後、個体レベルでの遺伝子機能解析に多大な貢献が期待できる。

Research Products

• [Publications] Hanazono,Y.: "Gene transfer into non-human primate hematopoietic stem cells : implications for gene therapy."Stem Cells. (in press).
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• [Publications] Shinjyo,T.: "Downregulation of Bim, a proapoptotic relative of Bcl-2, is a pivotal step in cytokine-initiated survival signaling in murine hematopoietic progenitors."Mol.Cell.Biol.. 21. 854-864 (2001)
• [Publications] Tarumoto,T.: "N (G)-monomethyl-L-arginine inhibits erythropoietin gene expression by stimulating GATA-2."Blood. 96. 1716-1722 (2000)
• [Publications] Kashii,Y.: "A member of Forkhead family transcription factor, FKHRLI, is one of the downstreammolecules of phosphatidylinositol 3-kinase-Akt activation pathway in erythropoietin signal transduction."Blood. 96. 941-949 (2000)
• [Publications] Urabe,M.: "Self amplification system for recombinant adeno-associated virus production."Biochem.Biophys.Res.Commun.. 276. 559-563 (2000)
• [Publications] Muroi,K.: "Sialyl-Tn- and neuron-specific enolase-positive minimally differentiated erythroleukemia."Intern.Med.. 39. 843-846 (2000)
• [Publications] Kawano,C.: "Engraftment syndrome after autologous peripheral blood stem cell transplantation with high numbers of peripheral blood stem cells followed by granulocyte colony-stimulating factor administration."Bone Marrow Transplant.. 25. 228-229 (2000)
• [Publications] Yoshida,K.: "Mediation by the protein-tyrosine kinase Tec of signaling between the B cell antigen receptor and Dok-1."J.Biol.Chem.. 275. 24945-24952 (2000)
• [Publications] Kume,A.: "Long-term tracking of murine hematopoietic cells transduced with a bicistronic retrovirus containing CD24 and EGFP genes."Gene Ther.. 7. 1193-1199 (2000)
• [Publications] Muroi,K.: "Autologous peripheral blood stem cell transplantation for adults with B-lineage acute lymphoblastic leukemia : a pilot study."Leukemia Lymphoma. 38. 103-111 (2000)
• [Publications] Urabe,M.: "DNA/calcium phosphate precipitates mixed with medium are stable and maintain high transduction efficiency."Anal.Biochem.. 278. 91-92 (2000)
• [Publications] Hibino,H.: "Haematopoietic progenitor cells from common marmoset as targets of gene transduction by retroviral and adenoviral vectors."Eur J Haematol. (in press).
• [Publications] Hase,H.: "Case Report : The avalability of TCR-V repertoires analysis with RT-PCR methods for the early detection of pulmonary relapsed T-cell malignancy after the autologous stem cell transplantation"Am J Hematol. 64. 124-127 (2000)
• [Publications] Watari,K.: "Identification of a melanoma antigen, PRAME, as a BCR/ABL-inducible gene."FEBS lett. 466. 367-371 (2000)
• [Publications] Tanabe T.: "Maxizyme, novel allosterically controllable ribozymes, can be designed to cleave various substrates."Biomacromolecules. 1. 108-117 (2000)
• [Publications] Machida,U.: "The effect of G-CSF administration in healthy donors before bone marrow harvesting."Br.J.Haematol,. 108. 747-753 (2000)
• [Publications] Tanabe T.: "Oncogene inactivation in a mouse model."Nature. 406. 473-474 (2000)
• [Publications] Wu MS: "MHC (Major histocompatibility Complex)-DRB genes and polymorphisms in common marmoset."J Mol Evol.. 51. 214-222 (2000)
• [Publications] Yusa,N.: "Transcription factor Sp3 activates the liver/bone/kidney-type alkaline phosphatase promoter in hematopoietic cells."J Leukoc Biol.. 68. 772-777 (2000)
• [Publications] Wang,Z.: "IL-12 synergizes with B7 1 enhance the antitumor immunity in C57BL/6 mice."Int J Mod Cancer Therapy. 3. 51-53 (2000)
• [Publications] Hirose,K.: "Involvement of MgcRacGAP in cytokinesis through associatins with mitotic spindle and midbody."J.Biol.Chem.. (in press).
• [Publications] Kawashima,T.: "STAT5 induces macrophage differentiation or M1 leukemia cells through activation of interleukin-6 production mediated by NF-kB p65."J.Immunol. (in press).
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• [Publications] Yamada,K.: "Constitutively active STAT5A and STAT5B in vitro and in vivo : mutation of STAT5 is not a frequent cause of leukemogenesis."Int.J.Hematol.. 71. 46-54 (2000)
• [Publications] Nosaka,T.: "Jaks and STATs in hematopoietic cells."Int.J.Hematol.. 71. 309-319 (2000)
• [Publications] Fujio,K.: "Molecular cloning of a novel type I cytokine receptor similar to the common gamma chain."Blood. 95. 2204-2211 (2000)
• [Publications] Misawa,K.: "Molecular cloning and characterization of a mouse homolog of human TNFSF 14, a member of the TNF superfamily."Cytogenetics and Cell Genetics. 89. 89-91 (2000)
• [Publications] Misawa,K.: "A method to identify cDNAs based on localization of green fluorescent protein fusion products. Proc."Natl.Acad.Sci.U.S.A.. 97. 3062-3066 (2000)
• [Publications] Morita,S.: "Plat-E : an efficient and stable system for transient packaging of retroviruses."Gene Therapy. 7. 1063-1066 (2000)
• [Publications] Tsuruga,H.: "Identification of novel membrane and secreted proteins upregulated during adipocyte differentiation."Biochem.Biophys.Res.Commun.. 272. 293-297 (2000)
• [Publications] Fujio,K.: "Functional reconstitution of Class-II MHC-restricted T cell immunity mediated by retroviral transfer of the α β TCR complex."J.Immunol.. 165. 528-533 (2000)
• [Publications] Kojima,T.: "Troy, a newly identified member of the tumor necrosis factor receptor superfamily, exhibits a homology with Edar and is expressed in embryonic skin and hair follicles."J.Biol.Chem.. 275. 20742-20747 (2000)
• [Publications] Ariyoshi,K.: "Constitutive activation of STAT5 by a point mutation in the SH2 domain."J.Biol.Chem.. 275. 24407-24413 (2000)
• [Publications] Kawashima,T.: "MgcRacGAP is involved in the control of growth and differentiation of hematopoietic cells."Blood. 96. 2116-2124 (2000)
• [Publications] Sugiyama,T.: "A novel low-density lipoprotein receptor-related protein mediating cellular uptake of apolipoprotein E-enriched b-VLDL in vitro."Biochemistry. 39. 15817-15825 (2000)
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• [Publications] Matsuoka,T.: "Prostaglandin D2 as a mediator of allergic asthma."Science. 287. 2013-2017 (2000)
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• [Publications] Agata,Y.: "Histone acetylation determines the developmentally regulated accessibility for T cell receptor-g gene recombination."J.Exp.Med.. (in press).
• [Publications] Honda,K.: "Molecular basis for hematopoietic/ mesenchymal interaction during initiation of Peyer's patch organogenesis."J.Exp.Med.. (in press).
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• [Publications] Ohishi K.: "Gaal and Gpi8p are components of a glycosylphosphatidylinositol (GPI) transamidase that mediate attachment of GPI to proteins."Mol.Biol.Cell. 11. 1523-1533 (2000)
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• [Publications] Sano,S.: "Two distinct signaling pathways in hair cycle induction : Stat 3-dependent and -independent pathways"Proc.Natl.Acad.Sci.USA. 97. 13824-13829 (2000)
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• [Publications] Hanazono,Y.: "Cell Therapy"Springer-Verlag, Tokyo. 11 (2000)
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• [Publications] Kinoshita T.: "Animal Model of PNH, PNH and GPI-linked proteins"Academic Press. 20 (2000)